This invention relates to medical and nutritional uses of extracts and products containing cocoa polyphenols including cocoa procyanidins.
Polyphenols are an incredibly diverse group of compounds (Ferriera et al., Diversity of Structure and Function in Oligomeric Flavanoids, Tetrahedron, 48:10, 1743-1803, 1992). They widely occur in a variety of plants, some of which enter into the food chain. In some cases they represent an important class of compounds for the human diet. Although some of the polyphenols are considered to be non-nutritive, interest in these compounds has arisen because of their possible beneficial effects on health.
For instance, quercetin (a flavonoid) has been shown to possess anticarcinogenic activity in experimental animal studies (Deshner et al., xe2x80x98Quercertin and Rutin as Inhibitors of Azoxymethanol-induced Colonic Neoplasiaxe2x80x99, Carcinogenesis, 7:1193-1196, 1991: and Kato et al., xe2x80x98Inhibition of 12-O-tetradecanoylphorbol-13-acetate Induced Tumor Promotion and Ornithine Decarboxylase Activity by Quercitin: Possible Involvement of lipoxygenase Inhibitionxe2x80x99, Carcinogenesis, 4, 1301-1305 1983). (+)-catechin and (xe2x88x92)-epicatechin (flavan-3-ols) have been shown in inhibit Leukemia virus reverse transcriptase activity (Chu et al., Inhibitory Effects of Flavonoids on Maloney Murine Leukemia Virus Reverse Transcriptase Activity, J. of Natural Products, 55:2, 179-183, 1992). Nobotanin (an oligomeric hydrolyzable tannin) has also been shown to possess anti-tumor activity (Okuda et al., xe2x80x98Molecular Structures and Pharmacological Activities of Polyphenolsxe2x80x94Oligomeric Hydrolyzable Tannins and Othersxe2x80x99xe2x80x94Presented at the XVIth International Conference of the Groupe Polyphenols, Lisbon, Portugal, Jul. 13-16, 1992). Statistical reports have also shown that stomach cancer mortality is significantly lower in the tea producing districts of Japan. Epigallocatechin gallate has been reported to be the pharmacologically active material in green tea that inhibits mouse skin tumors (Okuda et al, xe2x80x98Molecular Structures and Pharmacological Activities of Polyphenolsxe2x80x94Oligomeric Hydrolyzable Tannins and others. Presented at the XVIth International Conference of the Groupe polyphenols, Lisbon, Portugal, 1992). Osakabe et al. (JP 7274894 xe2x80x9cFood and Drink For Preventing Gastric Ulcersxe2x80x94Contains Antioxidation Substance Extracted from Cacao Beans using Hot Water or Ethanolxe2x80x9d October 1995; JP 7213251 xe2x80x9cMethod on Manufacturing an Antioxidation Substance and a Health Food or Drink Item Containing an Antioxidation Substancexe2x80x9d August 1995) have reported that the antioxidant properties of cocoa bean extract, thought to contain epicatechin and analogous compounds, are responsible for inhibiting formation of gastric ulcers in rats. Ellagic acid has also been shown to possess anticarcinogen activity in various animal tumor models (Boukharta et al., Efficacy of Ellagitannins and Ellagic Acid as Cancer Chemopreventive Agentsxe2x80x94Presented at the XVIth International Conference of the Groupe Polyphenols, Lisbon, Portugal, Jul. 13-16, 1992). Lastly, proanthocyanidin oligomers have been reported by the Kikkoman Corporation for use as antimutagens (xe2x80x98Antimutagenic Agent Containing Proanthocyanidin Oligomer Preferably Having Flavan-3-ol-diol Structurexe2x80x99 JP 04190774A, Jul. 7, 1992). Indeed, the area of phenolic compounds in foods and their modulation of tumor development in experimental animal models has been recently presented to the 202nd National Meeting of The American Chemical Society (Phenolic Compounds in Foods and Their Effects on Health II. Antioxidants and Cancer Prevention, Huang, M. -T., Ho, C. -T., and Lee, C. Y. editors, ACS Symposium Series 507, American Chemical Society, Washington, D.C. 1992).
It has been surprisingly discovered that cocoa extract, and compounds therefrom, have anti-tumor, anti-cancer or antineoplastic activity or, is an antioxidant composition or, inhibits DNA topoisomerase II enzyme activity or, is an antimicrobial or, is a cyclo-oxygenase and/or lipoxygenase modulator or, is a NO or NO-synthase modulator or, is a blood or in vivo glucose modulator.
Accordingly, the present invention provides a substantially pure cocoa extract and compounds therefrom. The extract or compounds preferably comprises polyphenol(s) such as polyphenol(s) enriched with cocoa procyanidin(s), such as polyphenols of at least one cocoa procyanidin selected from (xe2x88x92) epicatechin, (+) catechin, procyanidin B-2, procyanidin oligomers 2 through 12, preferably 2 through 4 or 4 through 12, more preferably 3 through 12, and most preferably 5 through 12, procyanidin B-5, procyanidin A-2 and procyanidin C-1.
The present invention also provides an anti-tumor, anti-cancer or antineoplastic or antioxidant or DNA topoisomerase II inhibitor, or antimicrobial, or cyclo-oxygenase and/or lipoxygenase modulator, or an NO or NO-synthase modulator, or blood or in vivo glucose modulator composition comprising a substantially pure cocoa extract or compound therefrom or synthetic cocoa polyphenol(s) such as polyphenol(s) enriched with procyanidin(s) and a suitable carrier, e.g., a pharmaceutically, veterinary or food science acceptable carrier. The extract or compound therefrom preferably comprises cocoa procyanidin(s). The cocoa extract or compounds therefrom is preferably obtained by a process comprising reducing cocoa beans to powder, defatting the powder and, extracting and purifying active compound(s) from the powder.
The present invention further comprehends a method for treating a patient in need of treatment with an anti-tumor, anti-cancer, or antineoplastic agent or an antioxidant, or a DNA topoisomerase II inhibitor, or antimicrobial, or cyclo-oxygenase and/or lipoxygenase modulator, or an NO or NO-synthase modulator, or blood or in vivo glucose modulator comprising administering to the patient a composition comprising an effective quantity of a substantially pure cocoa extract or compound therefrom or synthetic cocoa polyphenol(s) or procyanidin(s) and a carrier, e.g., a pharmaceutically, veterinary or food science acceptable carrier. The cocoa extract or compound therefrom can be cocoa procyanidin(s); and, is preferably obtained by reducing cocoa beans to powder, defatting the powder and, extracting and purifying active compound(s) from the powder.
Anti-cancer, anti-tumor or antineoplastic or, antioxidant, DNA topoisomerase II enzyme inhibiting, antimicrobial, cyclo-oxygenase and/or lipoxygenase modulator NO- or NO-synthase and blood or in vivo glucose modulating activities, or compositions containing the inventive cocoa polyphenols or procyanidins can be prepared in accordance with standard techniques well known to those skilled in the pharmaceutical or food science or veterinary art(s).
Such compositions can be administered to a subject or patient in need of such administration in dosages and by techniques well known to those skilled in the medical, nutritional or veterinary arts taking into consideration such factors as the age, sex, weight, and condition of the particular subject or patient, and the route of administration. The compositions can be co-administered or sequentially administered with other antineoplastic, anti-tumor or anti-cancer agents, antioxidants, DNA topoisomerase II enzyme inhibiting agents, or cyclo-oxygenase and/or lipoxygenase, blood or in vivo glucose or NO or NO-synthase modulating agents and/or with agents which reduce or alleviate ill effects of antineoplastic, anti-tumor, anti-cancer agents, antioxidants, DNA topoisomerase II enzyme inhibiting agents, cyclo-oxygenase and/or lipoxygenase, blood or in vivo glucose or NO or NO-synthase modulating agents; again, taking into consideration such factors as the age, sex, weight, and condition of the particular subject or patient, and, the route of administration.
Further, the invention also comprehends a kit wherein the active cocoa extract is provided. The kit can include a separate container containing a suitable carrier, diluent or excipient. The kit can also include an additional anti-cancer, anti-tumor or antineoplastic agent, antioxidant, DNA topoisomerase II enzyme inhibitor or antimicrobial, or cyclo-oxygenase and/or lipoxygenase, NO or NO-synthase or blood or in vivo glucose modulating agent and/or an agent which reduces or alleviates ill effects of antineoplastic, anti-tumor or anti-cancer agents, antioxidant, DNA topoisomerase II enzyme inhibitor or antimicrobial, or cyclo-oxygenase and/or lipoxygenase, NO or NO-synthase or blood or in vivo glucose modulating agents for co- or sequential-administration. The additional agent(s) can be provided in separate container(s) or in admixture with the active cocoa extract. Additionally, the kit can include instructions for mixing or combining ingredients and/or administration.
A cocoa polyphenol-containing composition, which is useful for modulating inflammatory pathways in a mammal, for maintaining vascular health, and as an antibacterial treatment, comprising a cocoa ingredient containing the cocoa polyphenols and optionally a carrier, diluent, or excepient. The composition is useful as a food, a dietary supplement, or a pharmaceutical. The food or dietary supplement can be a beverage or an elixir (i.e., ethanol extract), or a powder.
The cocoa polyphenols can be from ingredients or prepared synthetically. They can be present in cocoa ingredients. They can be extracted from cocoa beans, cocoa nibs, or cocoa ingredients such as chocolate liquor, partially defatted cocoa solids, and/or fully defatted cocoa solids.
The cocoa procyanidins are monomers and/or oligomers of epicatechin and catechin. The oligomers include dimers through octadecamers.
When extracted from cocoa beans, cocoa nibs, cocoa nib fractions, chocolate liquor, partially defatted cocoa solids, and/or fully defatted cocoa solids, a solvent which dissolves the cocoa polyphenols is used. Suitable solvents include water, methanol, ethanol, acetone, ethyl acetate, or mixtures thereof. Preferred solvents are mixtures of water and methanol or acetone. When water is used as the solvent, it is preferable if it is slightly acidified. In some cases the extract is purified, for example by removal of the caffeine and/or theobromine, and then further purified by gel permeation chromatography and/or high pressure liquid chromatography. During the high pressure liquid chromatography, the extract can be fractionated into monomeric and oligomeric fractions containing at least 50% by weight of the monomers or specific oligomers. When the fractions contain the monomers and lower oligomers (up to and including the tetramer), the fractions contain about 90 to 95% by weight of the particular oligomeric fraction.
Use of the above composition provides a method for modulating a mammalian inflammatory pathway by inhibiting COX activity, for modulating the production of eicosanoids and endothelin, for preventing diseases (such as bowel disease edema, arthritis, gingivitis or peridontitis) caused by chronic inflammation, for preventing vascular disease, for enhancing nitric oxide synthesis, for inhibiting LOX activity, for reducing vasoconstriction, for reducing platelet aggregation, for inhibiting monocyte adhesion, for inhibiting vascular smooth muscle proliferation associated with vascular disease, for reducing thrombosis, for reducing blood pressure, and for modulating oxidative stress to prevent associated inflammatory disorders and vascular diseases. Modulation of oxidative stress, for example, by preventing (LDL) oxidation, is another method of which the above composition can modulate mammalian inflammatory pathways and vascular function and disease.
The products comprising the cocoa polyphenol-containing composition are preferably in forms suitable for oral delivery, such as tablets, capsules, pills, concentrates, powders, liquids, or food ingredients, food additives or dietary supplements. The tablet may comprise an effective amount of the cocoa polyphenol-containing composition and optionally a carrier, such as sorbitol, lactose, cellulose, or dicalcium phosphate. The capsule may comprise a gelatin capsule containing a predetermined dosage of the cocoa polyphenol-containing composition. The oral delivery product may also comprise a dietary supplement nutrient such as dicalcium phosphate, magnesium stearate, calcium nitrate, vitamins, and minerals.
The compositions comprising the cocoa extract, sub-fractions thereof or mixtures thereof further comprise a liquid or a solid carrier suitable for use in foods, food supplements or pharmaceuticals. Such products include food and beverage products, in addition to capsule, tablet and pressed powder compositions. For the purposes of this application, the following definitions will enable a clearer understanding of what is disclosed and claimed:
As used herein a xe2x80x9cfoodxe2x80x9d is a material consisting essentially of protein, carbohydrate and/or fat, which is used in the body of an organism to sustain growth, repair and vital processes and to furnish energy. Foods may also contain supplementary substances such as minerals, vitamins and condiments. See Merriam-Webster""s Collegiate Dictionary, 10th Edition, 1993.
As used herein, a xe2x80x9cpharmaceuticalxe2x80x9d is a medicinal drug. See Merriam-Webster""s Collegiate Dictionary, 10th Edition, 1993.
As used herein, a xe2x80x9cFood Supplementxe2x80x9d is a product (other than tobacco) that is intended to supplement the diet that bears or contains the one or more of the following dietary ingredients: a vitamin, a mineral, an herb or other botanical, an amino acid, a dietary substance for use by man to supplement the diet by increasing the total daily intake, or a concentrate, metabolite, constituent, extract or combination of these ingredients. See Merriam-Webster""s Collegiate Dictionary, 10th Edition, 1993. As used on food labels, xe2x80x98supplementxe2x80x99 means that nutrients have been added in amounts greater than 50% above the U.S. RDA (xe2x80x9cUnderstanding Normal and Clinical Nutrition, Third Editionxe2x80x9d, Eds. Whitney, Cataldo and Rolfes, p. 525).
The compositions comprising the cocoa extracts, sub-fractions thereof, or mixtures thereof, are useful for inhibiting COX activity, inhibiting LOX activity, enhancing nitric oxide production, reducing vasoconstriction, reducing platelet aggregation, inhibiting monocyte adhesion, inhibiting excessive proliferation of vascular smooth muscle, reducing thrombosis, reducing blood pressure and modulating the production of eicosanoids and endothelin. Diseases, such as bowel disease, arthritis, edema, gingivitis and peridontitis, which are caused by chronic inflammation, are also prevented.